Dna oligonucleotides

The plasmids containing the p50 and p65 subunit genes of NF-κB were obtained from Synbio Technologies, Inc. Ni-NTA agarose was purchased from Qiagen Inc. DNA oligonucleotides were from IDT (Integrated DNA Technologies). Acrylamide, ammonium persulfate, N,N′-methylene-bis-Acrylamide, dithiothreitol, potassium glutamate, acetic acid, ammonium acetate, magnesium acetate, E. coli tRNA, isopropyl β-D-thiogalactopyranoside (IPTG), phospho(enol)pyruvate, CTP, UTP, ATP, GTP, cAMP, amino acids, rifampicin, sodium pyruvate, glutamine, formamide, calcium ionophore A23187, PMA, phenylmethanesulfonyl fluoride (PMSF), and Nonidet P40 were purchased from Sigma-Aldrich, while SDS and Tris were from Bio-Rad Laboratories. [³⁵S]-Met (1000 Ci/mmol, 10 μCi/μL) and [γ−³²P]-ATP (6 Ci/mmol, 10 μCi/μL) were obtained from PerkinElmer Inc. T4 polynucleotide kinase and T4 RNA ligase were from New England Biolabs Inc. RPMI 1640 medium, streptomycin, penicillin, and fetal bovine serum (FBS) were purchased from the American Type Culture Collection (ATCC). The human Jurkat leukemia T cell line was from ATCC (TIB-152).
Phosphorimager analysis was carried out with an Amersham Biosciences Storm 820 utilizing ImageQuant software (version 5.2, Molecular Dynamics). UV spectral measurements were determined on a PerkinElmer Lambda 20 UV/vis spectrometer.